Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 102, Issue 37, Pages 13081-13086Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0406877102
Keywords
super resolution; moire; resolution extension; saturation
Categories
Funding
- NIGMS NIH HHS [GM31627, R01 GM031627, GM25101, R01 GM025101] Funding Source: Medline
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Contrary to the well known diffraction limit, the fluorescence microscope is in principle capable of unlimited resolution. The necessary elements are spatially structured illumination light and a nonlinear dependence of the fluorescence emission rate on the illumination intensity. As an example of this concept, this article experimentally demonstrates saturated structured-illumination microscopy, a recently proposed method in which the nonlinearity arises from saturation of the excited state. This method can be used in a simple, wide-field (nonscanning) microscope, uses only a single, inexpensive laser, and requires no unusual photophysical properties of the fluorophore. The practical resolving power is determined by the signal-to-noise ratio, which in turn is limited by photobleaching. Experimental results show that a 2D point resolution of < 50 nm is possible on sufficiently bright and photostable samples.
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